PROJECT SUMMARY: FLOW CYTOMETRY SHARED RESOURCE The Flow Cytometry Shared Resource (FCSR) at Sylvester Comprehensive Cancer Center (Sylvester) is a state-of-the-art facility offering comprehensive analytical flow cytometry and high-speed cell sorting services that are pivotal to the successful progress of many research projects. The FCSR Director and staff work collectively to provide professional consultation, assay design, and research training as well as full cytometric analysis and cell sorting services to support research programs, publications, and grant applications. The FCSR?s six sophisticated fluorescence flow cytometry instruments (three cell analyzers and three cell sorters) facilitate a wide range of complex experiments examining the biological properties of both normal and cancer cells. The FCSR has the capability to provide simultaneous analysis of up to 19 different fluorescent parameters and two laser light-scatter parameters (forward and side scatter) for both cell surface and intracellular antigen detection in complex cell mixtures. The FCSR performs a variety of experimental analyses, such as complex immunophenotyping, DNA content and cell cycle analysis, cell signaling and activation analysis, and measurement of proliferation and apoptosis. Further, the FCSR provides fluorescence-activated cell sorting services, which allow investigators to purify both bulk and single cells at high purity and high speed, based upon complex multiparametric phenotyping. The three cell sorters have the potential to flow-sort up to 17 fluorescence parameters into bulk tubes or into multi-well plates for single cell flow cloning under Biosafety Level 2 conditions. The FCSR?s three highly experienced technicians perform cell sorting as well as continuous quality control assessment for all instruments. They also provide comprehensive user training in all aspects of flow cytometric experiments, including experimental design, fluorescent reagent panel selection, instrument setup and operation, data acquisition, and data analysis. Users are trained such that they are capable of operating the analysis cytometers themselves and then are permitted to do so independently during or outside regular operating hours. Users access the facility through an online instrument reservation and invoicing system. In 2017, the FCSR provided cytometry services to 41 Sylvester members (95% of whom were supported by peer-reviewed funding), which comprised 78% of total FCSR usage.